Antagonistic CD40 monoclonal antibodies and uses thereof
The disclosure provides for antibodies that bind CD40, including a humanized antibody. The antibodies bind CD40 and do not exhibit CD40 agonist activity. The antibodies may comprise a modified IgG1 Fc domain, and exhibit minimal activation of immature dendritic cells. Compositions comprising antibodies, methods of use for treatment of diseases involving CD40 activity, and use in the preparation of a medicament for treatment of a disease involving CD40 activity are provided.
1. A nucleic acid molecule encoding an isolated antibody or antigen binding portion thereof, that specifically binds to human CD40, wherein said antibody comprises a first polypeptide portion comprising a heavy chain variable region, and a second polypeptide portion comprising a light chain variable region, wherein:
said heavy chain variable region comprises a CDR1 comprising SEQ ID NO: 1, a CDR2 comprising SEQ ID NO: 2, a CDR3 comprising SEQ ID NO: 3; and
said light chain variable region comprises a CDR1 comprising SEQ ID NO: 7, a CDR2 comprising SEQ ID NO: 8, and a CDR3 comprising SEQ ID NO: 9.
2. An expression vector comprising the nucleic acid molecule of claim 1 .
3. A cell transformed with the nucleic acid of claim 1 .
4. A method of preparing an anti-human CD 40 antibody, or antigen binding portion thereof, comprising:
a) expressing the antibody, or antigen binding portion thereof, in the cell of claim 3 ; and
b) isolating the antibody, or antigen binding portion thereof, from the cell.
5. The nucleic acid molecule of claim 1 , wherein said antibody or antigen binding portion thereof antagonizes a CD40 activity.
6. The nucleic acid molecule of claim 1 , wherein said heavy chain variable region comprises a CDR1 consisting of SEQ ID NO: 1, a CDR2 consisting of SEQ ID NO: 2, a CDR3 consisting of SEQ ID NO: 3; and
said light chain variable region comprises a CDR 1 consisting of SEQ ID NO: 7, a CDR2 consisting of SEQ ID NO: 8, and a CDR3 consisting of SEQ ID NO: 9.
7. The nucleic acid molecule of claim 1 , wherein said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 4, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 10.
8. The nucleic acid molecule of claim 1 , wherein said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 4, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 16.
9. The nucleic acid molecule of claim 1 , wherein said first polypeptide portion comprises a human heavy chain constant region; and said second polypeptide portion comprises a human light chain constant region.
10. The nucleic acid molecule of claim 9 , wherein said human heavy chain constant region is a human IgG1 Fc domain comprising either
(1) a mutation at Kabat position 238 that reduces binding to Fc-gamma-receptors (FcγRs), wherein proline 238 (P238) is mutated to one of the residues selected from the group consisting of: lysine, serine, alanine, arginine, and tryptophan, and wherein the antibody or antigen binding portion thereof has reduced FγR binding; or
(2) an alanine substituted at Kabat position 297.
11. The nucleic acid molecule of claim 9 , comprising a human IgG1 Fc domain comprising a mutation at Kabat position 238 that reduces binding to Fc-gamma-receptors (FcγRs), wherein proline 238 (P238) is mutated to one of the residues selected from the group consisting of: lysine, serine, alanine, arginine, and tryptophan, and wherein the antibody or antigen binding portion has reduced FcγR binding.
12. The nucleic acid molecule of claim 11 , wherein P238 is mutated to lysine.
13. The nucleic acid molecule of claim 12 , wherein the Fc domain comprises an amino acid sequence selected from: SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, or SEQ ID NO: 29.
14. The nucleic acid molecule of claim 10 , wherein said heavy chain variable region comprises a CDR1 consisting of SEQ ID NO: 1, a CDR2 consisting of SEQ ID NO: 2, a CDR3 consisting of SEQ ID NO: 3; and
said light chain variable region comprises a CDR1 consisting of SEQ ID NO: 7, a CDR2 consisting of SEQ ID NO: 8, and a CDR3 consisting of SEQ ID NO: 9.
15. The nucleic acid molecule of claim 14 , wherein the human IgG1 Fc domain comprises the amino acid sequence of SEQ ID NO: 22 or SEQ ID NO: 23.
16. The nucleic acid molecule of claim 1 , wherein the first polypeptide portion comprises or consists of an amino acid sequence selected from the group consisting of:
SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 30, and SEQ ID NO: 31; and
the second polypeptide portion comprises or consists of the amino acid sequence of SEQ ID NO: 11.
17. The nucleic acid molecule of claim 1 , wherein the first polypeptide portion comprises an amino acid sequence of SEQ ID NO: 5; and
the second polypeptide portion comprises the amino acid sequence of SEQ ID NO: 11.
18. The nucleic acid molecule of claim 1 , wherein the first polypeptide portion consists of an amino acid sequence of SEQ ID NO: 5; and
the second polypeptide portion consists of the amino acid sequence of SEQ ID NO: 11.
19. The nucleic acid molecule of claim 1 , wherein the isolated antibody or antigen binding portion thereof is humanized.
20. The nucleic acid molecule of claim 1 , wherein the antigen binding portion is an scFv-Fc.
21. The nucleic acid molecule of claim 1 , wherein the antibody or antigen-binding portion thereof is linked to a therapeutic agent.
22. The nucleic acid molecule of claim 1 , wherein the antibody or antigen-binding portion thereof is linked to a second functional moiety having a different binding specificity than said antibody or antigen binding portion thereof.
23. The nucleic acid molecule of claim 1 , further comprising an additional moiety.